摘要目的 分析AD与其部分血管性危险因素的关系,探讨丹参酮ⅡA(TanⅡA)对AD合并慢性脑缺血小鼠模型的干预作用及相关机制.方法 收集苏州大学附属第一医院神经内科自1975年至2009年收治的849例痴呆患者(确诊AD 549例,非AD 300例)的临床资料,单因素分析AD与血管性因素的相关性.Tg+小鼠按随机数字表法分为5组(假手术组、模型组、模型+TanⅡA低剂量组、模型+TanⅡA中剂量组、模型+TanⅡA高剂量组)或3组[假手术组、模型组、模型+TanⅡA 10 mg/(kg.d)组],分别建立AD合并慢性脑缺血小鼠模型并给予TanⅡA干预.Morris水迷宫实验检测小鼠学习记忆能力的变化,ELISA法检测血浆血管内皮生长因子(VEGF)水平,RT-PCR检测脑组织Aβ前体蛋白(APP)、VEGF、VEGFR-1 mRNA含量,Western blotting检测脑组织APP、Aβ、VEGF、VEGFR-1蛋白表达.另在细胞水平观察TanⅡA对人脐静脉内皮细胞(HUVEC)成管的影响及VEGFR-1 mRNA和蛋白的表达变化.结果 AD与高血压病、糖尿病、冠心病、脑血管病、高脂血症等血管性危险因素均呈显著正相关关系(P＜0.05),而与非血管性因素肺炎则无相关关系.与模型组比较,模型+TanⅡA中剂量组、模型+Tan ⅡA高剂量组的寻台时间及游泳距离明显缩短,避暗潜伏期明显延长、错误次数减少,血浆VEGF表达明显下调,差异有统计学意义(P＜0.05).与模型组比较,模型+Tan ⅡA 10 mg/(kg.d)组的中位生存时间延长约24d,APP、VEGF mRNA表达明显下调,VEGFR-1 mRNA表达明显上调,APP、Aβ、VEGF蛋白表达明显下调,VEGFR-1蛋白表达明显上调,差异有统计学意义(P＜0.05).低氧+TanⅡ A 5μg/mL组HUVEC细胞毛细管形态完整且密度升高,VEGFR-1 mRNA和蛋白表达较低氧组明显上调,差异有统计学意义(P＜0.05)结论 AD发病与其血管性因素密切相关.TanⅡA可能通过上调VEGFR-1表达促进毛细管结构的完整性,从而改善模型小鼠痴呆症状,提示干预血管性因素可能防治AD.

abstractsObjective To evaluate the relationship between Alzheimer′s disease(AD)and its partial vascular risk factors,and investigate the effect of tanshinone ⅡA(Tan ⅡA)on experimental models of AD combined with chronic cerebral ischemia and its potential mechanism.Methods Eight hundred and forty-nine patients with dementia(549 having diagnosis of AD and 300 without AD),admitted to our hospital from 1975 to 2009,were collected in our study; univariate analysis was performed on the relation between AD and vascular risk factors.Besides that,Tg+ mice were employed in our study and randomly divided into 5 groups,namely,sham-operated group,vehicle group,low Tan ⅡA treatment group,medium Tan ⅡA treatment group and high Tan ⅡA treatment group; or these mice were randomly divided into sham-operated group,vehicle group,Tan ⅡA treatment group(treated with 10 mg/kg daily).Mouse models of AD and chronic cerebral ischemia were established,and Tan ⅡA treatment was given to the Tan ⅡA treatment group.The relationship between AD and vascular factors was assessed by means of analyzing the clinicopathological data of AD cohort.The changes of learning and memory abilities in the mouse models were detected by Morris water maze test.Enzyme-linked immuno sorbent assay(ELISA)was employed to detect the level of VEGF; the protein expressions of betaA4-amyloid precursor protein (APP),VEGF and VEGFR-1 were determined by Western blotting,and the mRNA expressions of APP,VEGF and VEGFR-1 were observed by quantitative RT PCR.The effect of Tan ⅡA on canaliculization of human umbilical vein endothelial cells(HUVECs)was also investigated fiom cellular level.Results AD was significantly positively correlated with such vascular risk factors as hypertension,diabetes mellitus,coronary disease,cerebrovascular disease and hyperlipemia(P＜0.05),while no correlation was noted between AD and pneumonia.The mice of the medium Tan ⅡA treatment group and high Tan ⅡA treatment group had obviously shortened times of searching the platform and swimming distance,prolonged latency of avoiding darkness,decreased frequency of wrong behaviors,and decreased level of VEGF as compared with the vehicle group(P＜0.05).The life span in mice of the Tan ⅡA treatment group (treated with 10 mg/kg daily)was prolonged for approximately 24 d as compared with that in the vehicle group; the expressions of APP and VEGF were down-regulated and that of VEGFR-1 in mice of the Tan ⅡA treatment group(treated with 10 mg/kg daily)was up-regulated as compared with those in the vehicle group(P＜0.05).The canaliculization of HUVECs was enhanced after incubation with Tan ⅡA for 48 h,followed by increase of VEGFR-1 expression.Conclusion AD is significantly correlated with its vascular factors.Tan ⅡA could improve the learning and memory abilities of dementia mouse through up-regnlation of VEGFR-1 expression and promotion of vascular integrity,indicating the crucial role of vascular factors in treatment of AD.